Neonatal sepsis causes over one million death per year worldwide and 5% to 60% of infants treated with antibiotics die despite the treatment, with higher rates occurring in low income countries. Rapid diagnosis of sepsis in NICUs (Neonatal Intensive Care Units) is crucial for implementing a timely treatment. Because sepsis is a life-threatening medical emergency a large proportion of infants receive treatment with potent systemic antibiotics that can adversely lead to destruction of the infant's normal gastrointestinal flora and the risk of becoming colonized with drug-resistant microorganisms. Blood culturing is used as the gold standard for neonatal diagnosis of sepsis, however the samples are returned in 2 to 3 days which often does not provide timely results for neonatal sepsis cases. Typical neonatal patients are weighing 1,000 grams or less and their total blood volume may be as low as ~ 50 mL, thus the iatrogenic blood loss for premature babies may represent up to 12% - 31% of total blood loss due to phlebotomy. New point-of-care methods with faster detection time and using smaller amounts of sample are urgently needed. Molecular assays based on DNA specificity for sepsis pathogens represent a promising diagnostic tool for early identification of bacteremia. However, today 90% of hospitals in USA cannot perform DNA analysis and samples are sent to the centralized reference labs with the response time of 2-3 days. The current point-of-care diagnostics are mostly based on the real time PCR methods that are limited by the number of colors for detecting multiple organisms (maximum 4-6) or by the high price of the instrument or assay cost. This project introduces a new point-of-care (POC) rapid method for DNA/RNA diagnostics that will approach total assay turn-around time of ~15 min and that is capable of using very small sample volumes, as low as 2 - 200 microliters, compatible with the neonatal sepsis detection. Multiplexed identification of the neonatal sepsis pathogens is enable using the microarray approach. In addition, a number of innovations are offered in the project that will bring a 5 fold decrease in the cost of instrumentation and cartridges compared to the commercialized platforms. The platform also offers a unique new qualitative parameter in DNA/RNA diagnostics that will enable integration of data for both the length of the nucleic acids as well as it will preserve the strand specificity, e.g., important in alternative splicing mechanisms. The assay development in the Phase I will focus on representative neonatal sepsis pathogens and will establish the basic platform, and cartridge prototypes and will provide a thorough validation of the assays developed using de- identified archived clinical samples.